LOCUS       sequence-327123-        8889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of PmcCAST TnsAB, TnsC and TnsD proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8889)
  AUTHORS   Saito M, Ladha A, Strecker J, Faure G, Neumann E, Altae-Tran H, 
            Macrae RK, Zhang F
  TITLE     Dual modes of CRISPR-associated transposon homing.
  JOURNAL   Cell. 2021 Apr 29;184(9):2441-2453.e18. doi: 
            10.1016/j.cell.2021.03.006. Epub 2021 Mar 25.
  PUBMED    33770501
REFERENCE   2  (bases 1 to 8889)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2021.03.006"; journalName: "Cell"; date: 
            "2021-04-29- 29"; volume: "184"; issue: "9"; pages: "2441-2453.e18"
COMMENT     Sequence Label: pMS26: pTns(PmcCAST)_Delta-TniQ
FEATURES             Location/Qualifiers
     source          1..8889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        67..85
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    86..110
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        5434..5452
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5453..5477
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             5586..5630
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(5668..5686)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5682..5729
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5850..6641)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(6642..6733)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6781..7519)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(7729..8811)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     8773..8792
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8812..8889)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aagcggctat ttaacgaccc tcgggatctc gacgctctcc cttatgcgac tcctgcatta
       61 ggaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctgtagaaa
      121 taattttgtt taactttaat aaggagatat accatgaaag agcaatttga acaatgggtt
      181 gctcgccttc aaatatccga ggcagggcaa aaaattattg agaaagttcg ctcatctgag
      241 ccatcccgtc gcgttggtgg cggtagcaag aatgtttcgg gacgctaccc tagtcgcaag
      301 atgggagtca cgattcaatt tgagtctcat cgggttgaac tgccaattat ctatcagtta
      361 gaacatgata aagatgttct agaattctac gatcaaccac cgcaaattaa acttgactat
      421 caagcaagta acggacgtag attaggcgtt ttacacaccc cagatttatt tgtcatccga
      481 accaattcgg ctggttggga agagtgtaaa actgagcagg atttgaaaaa actagcagag
      541 aaaaacccca atcgttactt ttactctgaa gaaaataatc aatggcattc cttgccagga
      601 gaagtttatg ctaaccagtt tgggctttat caccgcattc attcagatcg cgaaatcaat
      661 tgggttctgc aacgtaattt gcaattttta gaagattatt atcgctcaga atctctagta
      721 gtggaagaag cgatcgctca atcattactt gcaattgtgt catctcaacc aggaatcact
      781 ttagcagaat tactcaatcg gtcaacagga gctaaatctg acgatatcta caccttaatt
      841 attcaagagc aaatctatat tgatttaaat gctacaccac tagctgaacc agaaagatgt
      901 ctaatcttcc gtgacgagca aactgctagt gcttatagat taatggttga gcaaccaagt
      961 gtaagtcttc cggctatctc tcctgtagtc aatattgtta ctggagcaac tgttagttgg
     1021 gacggcaaag gtttaaatat catccatgtt ggagaaacag aggttatact tggcgcagaa
     1081 aataatcagt taattgaact caaaaaagct atatttgaga atctggttcg ccaaggaaaa
     1141 atcacaagtc tccaaacacc agaaaaaaca gctattagca cagaatcttg gcagcgattt
     1201 catcaagcta gtcctgaaga ccaagcagaa gcccttgaac gctacaaaac tatcgaacct
     1261 tacttaaacg gacatcctcc agaaaacgaa acaataccag cccgtaccat tcgtcactgg
     1321 aaagctaaat atttaattgc aattcaaaaa tacggttgcg gctacattgg gcttttgtcc
     1381 catcgcagtg tcaaaggtaa tcgacagcgc aagctgcccg aagacacctt agcaattatg
     1441 gagaagttta ttttagaaga ctatgaaact cttaaacaaa aaagaatgtg ggaggttcac
     1501 gcggctttag ttcgagcttg cgaagaagct ggtgtaattg ctccgagcta taaagcattt
     1561 accaaagaag tacaacgccg gactggttac gaacagacta aaaaacgcca aggtcgtcgt
     1621 gccgcttatc agcatgaatc cttctattgg gagcttgcta taactacccc aagacacggc
     1681 gatcgccctt ttgaaattgg gcatatagat catactgagt tagatgtaga gctagtttgc
     1741 tctgatacag gtcgcaatct tggcagacct tgggcaacat ttctcgtaga tgcttattca
     1801 cggcgattgc tggcagttta cctaacgttt gactcaccct catatcgttc ttgcttaatg
     1861 gtgctaagaa tttgtgtcaa gcgtcatggc aggcttcccc aaattgtagt ggtagataac
     1921 ggtgcagagt ttcacagcgt ttactttgaa acactactag ctacttttga atgtacaaaa
     1981 aagcaaagac ccccagccaa agctcgtttt ggttcggttt gcgaacgcct atttggaact
     2041 tccaatactc aatttgtcca taacttactt ggtaatacac aaatcactcg taatgtccgc
     2101 caagtcacta agtcagttaa ccctaaaaat ctagctgtat ggacacttgg tttactttat
     2161 gaatatcttt gtgcttgggc ttacgaagtt tatgatacag acgaacatcc ggcacttttc
     2221 cagagtccga gggatgcgtt tgctgcgggt atggcgattg gaggcagtcg ggttcaccgc
     2281 atgattcctt acgatgaaaa ctttcagatt cttaccctac ctacaacctc agagggaaaa
     2341 gcaaaagttc aagttggacg cggcataaaa attaactaca tatactactg gtctaatagt
     2401 ttccgcgatc ctcaaattga aaatacctct gtgcagataa gatacgaccc gtttaatatt
     2461 ggcatcgctt acgcctttgt tcgaggacag tgggtacagt gtatctctca atattatgca
     2521 gaactccaag gtcgttcaga aaaagaactg aagttagcta gtatcgagtt acgcaaacac
     2581 tattcaaatc atacacaaca atctaaagtt tcggctaaga atttagcaga atttttagca
     2641 tcggtagaag ctcaagaagc tctattagaa cagcgttcct atgatgcaga agttaaggaa
     2701 gtctttaggg taatcgaggg tggaaaagcg acaacaagcc gtaacgaaga accaaaactt
     2761 attcaagtta cttttgccaa taccgatttt caagcggatg aagatgaagc gatcgtccca
     2821 gaaacacttg tagtgtatga ggagttttaa ctatgacaaa atctactgga tttccgcttg
     2881 aactcctcac acggcctgct acagaaaggc ttgcttactt tgaaaattac accgtagcgc
     2941 atcctcgact taaagaagtc tatgaaattc tcatgcgaac gattgccgaa ccagcaggag
     3001 cttcatttat ctttgtttac ggtgctagtg gtgttggtaa aacaactttg cgactccgcg
     3061 ttgagcagaa attaacagaa ctagcattac ctaaattgga aagcgatcgc gctcgtgttc
     3121 ctgttgttgg tattgaagct atagcgccag aatcgagata cttcaactgg aaagaatact
     3181 acacacgcgc cttaattact ttggaagaac cattaatcga tcacaagttt gattacggtg
     3241 tgcgtggtat ctctcgcgat aattttggca aaattaatgt tgagtcgaaa gtagttgcac
     3301 ctgctctacg cagagcttta gaaaatgctc tcatccatcg tcatccagac gtattttttg
     3361 tagatgaagc acaacatttt ggtaaggtgg caagcggata caaactacaa gatcaattag
     3421 actgtctcaa atctttagca aatatgacag gcatcttgca ctgtctatta ggaacatacg
     3481 aactacttac attccgtaat ttgagcggtc aactctcacg tcgtagtgtt gatattcatt
     3541 tccgccgtta ttgtgctgat agcccagaag atgtacaggc ttttaagagt gtgttgttaa
     3601 cttttcaaca gcatctgcca ctagcagaaa ctccaaattt agtcgatcat tgggaatatt
     3661 tttacgaacg tactttagga tgtattggca cactgaaaga ttggctgaag cgtgttctta
     3721 gtgatgcttt ggatcgagaa gcaacaacaa tcactctcaa agatttacaa aaacgtgctt
     3781 tgtcggttgc tcagtgtcag aaaatgttca aggaaattca agagggtgaa cgacaattat
     3841 ctgaaacaga agcagatgtg caaaacttac gctctgcatt aggacttggc gcaaaaccaa
     3901 ttgtgctacc cgaagaaacc ccaaaaacta ctcgtccgcc aggaaaggtt ggcaaacgca
     3961 aacctaaaag agatcccatc ggagtgcagc aagatgttag ctagtgtatt agaaacatac
     4021 gaatcttgga atttaaaaaa gccattaatt ccacaacgta gtcgcctcta tcaacctcaa
     4081 cctgtaggta ttgggactcc ttacatagag agtttaacag gttatatcac acggatagca
     4141 gagttgcatg gagttcttcc aggagtcttg atgactagag aaattgctcc gcttgtcaat
     4201 aaaatatatt tccaaaatgg agctaatcga ggttttcgtg aaatattcaa ccgtagtcaa
     4261 gcgttaaatg gcatgggaga gatggcagca gatttagttc aagtgttgca aaaattgacg
     4321 cttagagatg acttgcgttt tttaaccatg ctgttttggt ctaacatttt gactccacgc
     4381 aatttatttc gtaccagaaa agcttggtgt ccaatttgtt atcaagagcg gcatcaaaat
     4441 gggctggtag tctatgagca attgctttgg acaattaacc taattacaat ttgcccgcag
     4501 catcaaaaac ccttggttga actttgccct cactgtaatc atgaatcacc tttgttaaac
     4561 tggcgctcaa gacctggtta ctgttcaaag tgtggagagt ggttaggcgc aaatcaatgc
     4621 cttaagacat ttacagatgg tgaaggatca ataaaattac agttggagtg gcagtattgg
     4681 actgccaatg ttgtaggaga gctaatttta gcttctcaat gctttgaatc tgccccatca
     4741 aaagaaaata ttacaaaatc attaaatata gttattgata aagttgcgga aaataatgca
     4801 gctgcatttt ctcgtcttat tggagtacca aaaaatagtt tgtggatgtg gcagtctact
     4861 aaaaccttgc cagagctaaa tacacttctg aaaatttgtt acgaattaga gatatcgttg
     4921 gtagaatttc ttactccaaa aaatctaatt actaaatctt ttacaaaaat ctcgcagaag
     4981 cacctccaac tttctcgcac accaagggta tctccaaaat ctttcgacca atatcaagta
     5041 aaagatgctt tattagcaat acttgcaggt aatgaagaac caccaccaac aatggaagaa
     5101 gttggaaagc gtttaggtca tcacaacaga actatttcta gacactttcc agatttgtgt
     5161 agtgctattt ctgctaaatg tcgtaattac aataaagctt gccgtctcaa aagcatagaa
     5221 aaactttgca gcgaagttag agagattgtc cttagcctta atgctcaggg tgtgtatccc
     5281 acagaaggtc gtgtttgtga attaatgcca aatccggggt gttttagata taaacaagtc
     5341 cgcgctgctt ttaatgatgc acgacgtgag tttggtttgt aacttaagtc gaacagaaag
     5401 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
     5461 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
     5521 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
     5581 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
     5641 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     5701 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
     5761 tcacactgct tccggtagtg ccctgaaccg acgaccgggt catcgtggcc ggatcttgcg
     5821 gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct
     5881 ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct
     5941 tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc
     6001 tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac
     6061 caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag
     6121 ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca
     6181 aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc
     6241 aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg
     6301 cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg
     6361 tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc
     6421 gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc
     6481 accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg
     6541 tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct
     6601 gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct ttttcaatat
     6661 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6721 aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag actgcggcgg
     6781 gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg
     6841 tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca
     6901 gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg
     6961 aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt
     7021 cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt
     7081 tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt
     7141 gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg
     7201 cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact
     7261 ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg
     7321 ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc
     7381 tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc
     7441 acctccccag gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa aaaaaggatc
     7501 tcaagaagat cctttgatct tttctactga accgctctag atttcagtgc aatttatctc
     7561 ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc
     7621 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     7681 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     7741 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7801 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     7861 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     7921 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     7981 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     8041 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     8101 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     8161 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     8221 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     8281 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     8341 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     8401 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     8461 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     8521 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     8581 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     8641 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     8701 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     8761 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     8821 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     8881 gatggtgtc
//
